The present invention relates to monoclonal antibodies (mAb) that specifically react with the HLA-DR antigen and induce apoptosis in HLA-DR expressing cells. In particular, the invention relates to the use of such monoclonal antibodies in the treatment of cancers involving HLA-DR positive cells, and to pharmaceutical compositions containing anti-HLA-DR antibodies.
Babbitt, et al., Nature 317:359-361 (1985).
Borner, et al., J. Immunol. 147(1):86-95 (1991).
Bradley, L. M., SELECTED METHODS IN CELLULAR IMMUNOLOGY 153-174, Mishell and Shiigi, Eds., W. H. Freeman and Co., New York (1980).
Brodeur, et al., MONOCLONAL ANTIBODY PRODUCTION TECHNIQUES AND APPLICATIONS, Marcel Dekker, Inc., New York, pp. 51-63 (1987).
Cohen and Duke, Annu. Rev. Immunol. 10:267-293 (1992).
Cole, et al., MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, p. 77 (1985).
David, et al., Biochemistry 13:1014 (1974).
Fu and Karr, Human Immunol. 39:253-260 (1994).
Goding, MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE, Academic Press, 59-103 (1986).
Harlow and Lane, ANTIBODIES: A LABORATORY MANUAL, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1988).
Holland, et al., Cancer Med. 2:2697-2828 (1996).
Hoogenboom and Winter, J. Mol. Biol. 227:381 (1991).
Hunter, et al., Nature 144:945 (1962).
Jones, et al., Nature 321:522-525 (1986).
Kohler and Milstein, Nature 256:495 (1975).
Kozbor, J. Immunol. 133:3001 (1984).
Lampson and Levy, J. Immunol. 125:293-299 (1980).
Marks, et al., J. Mol. Biol. 222:581 (1991).
Mollick, et al., J. Immunol. 146:463-468 (1991).
Munson and Pollard, Anal. Biochem. 107:220 (1980).
Newell, et al., Proc. Natl. Acad. Sci., 90(22):10459-10463 (1993).
Nygren, J. Histochem. and Cytochem. 30:407 (1982).
Otten and Yokoyama, CURRENT PROTOCOLS IN IMMUNOLOGY, 5.4.1-5.4.19, Ed., Greene and Wiley, N.Y. (1997).
Pain, et al., J. Immunol. Meth. 40:219 (1981).
Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).
Riechmann, et al., Nature 332:323-329 (1988).
Ritts, et al., Int. J. Cancer 31:133-141 (1983).
Stocker, et al., Research Disclosure 217:155-157 (1982).
Swat, et al., J. Immunol. Methods 137:79-87 (1991).
Truman, el al., Blood 89(6):1996-2007 (1997).
Truman, el al., Int. Immunol. 6(6):887-896 (1994).
Verhoeyen, et al., Science 239:1534-1536 (1988).
Vermes, et al., J. Immunol. Meth. 184:39-51 (1995).
Vidovic, et al., Eur. J. Immunol. 25:3349-3355 (1995).
Vidovic and Toral, Cancer Lett. 128:127-135 (1998).
Vitale, et al., Histochemistry 100:223-229 (1993).
Wyllie, et al., J. Pathol. 142:67-77 (1984).
Zola, MONOCLONAL ANTIBODIES: A MANUAL OF TECHNIQUES, CRC Press, Inc. pp. 147-158 (1987).
Class II major histocompatibility complex (MHC) molecules, constitutively expressed on normal antigen presenting cells (APC), are responsible for the presentation of antigen-derived peptides to CD4+ helper T (Th) cells. (Babbitt, et al., 1985; Truman, et al., 1997). Signaling via these molecules initiates the generation of second messengers leading to programmed cell death (PCD or apoptosis) of activated B lymphocytes. Besides antigen presentation, class II molecules transduce signals that can modulate cell growth and certain class II MHC-specific mAb have been shown to induce apoptosis of cancer cells (Newell, et al., 1993). The practical utility of this observation in cancer therapy has been hampered by the intrinsic lack of selectivity, in that the class II MHC-specific antibodies that have been shown to induce apoptosis of cancer cells, also interfere with normal Th cell functions. (Vidovic, et al., 1995). More specifically, the presently available apoptosis-inducing class II-specific mAb recognize epitopes located on the first protein domains of the HLA-DR heterodimer, in apparent close proximity to the peptide-binding site, and these mAbs interfere with antigen presentation, causing a potent in vitro and in vivo inhibition of Th responses (Vidovic, et al., 1995).
Hence, the main problem in using the currently available anti-HLA-DR antibodies for the treatment of cancers involving HLA-DR positive cells is the potential for side effects such as immunosuppression of HLA-DR mediated immune responses based on the lack of definitive specificity of the antibodies for the apoptogenic epitope, and as a result such anti-HLA-DR antibodies may not find practical utility in therapeutic applications.
One approach to overcoming these problems is to administer an anti-Class II (anti-HLA-DR) antibody specifically reactive with tumor cells which can trigger apoptosis in such cells and which does not have immunosuppressive properties associated with the binding to HLA-DR-expressing cells.
Accordingly, it is an object of the invention to provide a composition for in vivo administration, comprising a monoclonal antibody which specifically binds to HLA-DR-expressing tumor cells and triggers apoptosis of the tumor cells to which it binds.
The present invention is based, in part, on the discovery that antibodies which specifically react with human major histocompatibility complex (MHC) class II can induce apoptosis of cells expressing HLA-DR molecules on their surface.
The antibodies of the present invention are highly specific in that the monoclonal antibodies affect neither the viability nor function of non-neoplastic HLA-DR positive cells.
In one aspect, the invention includes Fab fragments of monoclonal antibodies specific for HLA-DR anchored to a solid support.
The apoptosis-inducing effect of such monoclonal antibodies is associated with a cross-linking of HLA-DR, and monovalent Fab fragments cannot mediate cytotoxicity (Vidovic and Toral, 1998).
According to an important feature of the present invention, the tumoricidal effects of anti-class II MHC mAb can be achieved without simultaneous suppression of class II-dependent immune responses, although both properties are associated with the mAb recognizing an epitope in the first protein domains of HLA-DR.
As outlined herein, the anti HLA-DR antibodies of the present invention specifically bind to the first domains of HLA-DR molecules which are expressed by a variety of types of cancer cells, including, but not limited to B cell cancers. As further outlined herein, the anti-HLA-DR antibodies exert a triggering effect on apoptosis which is specific to HLA-DR positive tumor cells.
An important practical implication of this work is that a mAb, designated Danton; and produced by a hybridoma cell line, also designated Danton; may be effective for the selective antibody-based therapy of HLA class II positive neoplasms, including, but not limited to, blood cell neoplasms, e.g. plasmacytoma/multiple myeloma, Hodgkin""s and non-Hodgkin""s lymphomas and B cell leukemias. In vitro studies indicate that the Danton mAb does not interfere with normal Th function, therefore, therapy with the Danton mAb should not affect the subject""s normal HLA-DR-expressing cells. Accordingly, it would be reasonable to expect fewer side effects than with the currently available therapeutic agents.
These and other objects and features of the invention will become more fully apparent when the following detailed description of the invention is read in conjunction with the accompanying drawings.